| Abstract [eng] |
Engineering Recombinant Fusion Cytokines and Investigation of Their Activity The aim of this work was to fuse cytokines to engineer a recombinant chimeric protein GCSF/Lα/SCF and investigate the activity of this and several other proteins. It was done using total internal reflection ellipsometry (TIRE) which required immunosensors that were also created. The interactions between GCSF/Lα/SCF, SCF/Lα/GCSF and their receptors GCSF-R and c-kit were measured. Also, GCSF/Lα/GCSF binding to GCSF-R and SCF monomer binding to c-kit were measured. Mathematical models were applied to calculate association and dissociation rate constants for evaluation of interactions between receptors and ligands. GCSF/Lα/SCF was successfully engineered and immunosensors created for measuring the binding of the proteins. It was found that GCSF/Lα/SCF is able to bind to c-kit the fastest when compared to other proteins, but it does not interanct with GCSF-R. SCF/Lα/GCSF interacts with both receptors – it binds to GCSF-R slightly slower than homodimer and faster to c-kit than SCF monomer. Also, it dissociates slower than SCF. |
